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Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

  • Authors:
    • Chao Li
    • Lei Yang
    • Yudong Zhang
    • Baocheng Zhao
    • Hao Qu
  • View Affiliations / Copyright

    Affiliations: Division of Gastrointestinal Surgery, Department of General Surgery, Beijing Chao‑Yang Hospital, Capital Medical University, Beijing 100020, P.R. China, Medical Research Center, Beijing Chao‑Yang Hospital, Capital Medical University, Beijing 100020, P.R. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 81
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    Published online on: February 25, 2026
       https://doi.org/10.3892/or.2026.9086
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Abstract

Hepatocellular carcinoma (HCC) is an aggressive liver malignancy, the molecular mechanisms underlying the progression of which are not fully understood. As a component of the chaperonin‑containing tailless complex polypeptide 1 (TCP1) ring complex, chaperonin‑containing TCP1 subunit 2 (CCT2) participates in the development of numerous types of liver disease. However, the potential role of CCT2 in regulating HCC malignant behaviors remains unclear. In the present study, bioinformatics analysis of patients with HCC from public datasets (The Cancer Genome Atlas‑Liver HCC, International Cancer Genome Consortium‑Liver Cancer‑Riken‑Japan and OEP00000321) demonstrated that CCT2 expression was upregulated in HCC tissue. High expression of CCT2 was also associated with an unfavorable overall survival prognosis. CCT2 knockdown was shown to inhibit the proliferation, migration, invasion and stemness and promote the apoptosis of HCC cells in vitro, as evidenced by EdU, colony formation, flow cytometry, caspase‑3/7 activity, gap closure, Transwell and tumor‑sphere formation assays. Consistently, knocking down CCT2 also suppressed the subcutaneous tumor proliferation and hematogenous lung metastasis of the human HCC HCCLM3 cells in vivo. Furthermore, downregulation of CCT2 decreased the phosphorylation of STAT3, as well as the expression of myeloid cell leukemia sequence 1, matrix metalloproteinase 2 and SRY‑box transcription factor 2 in vitro and in vivo. However, IL‑6 treatment rescued the levels of phosphorylated STAT3 and counteracted the inhibitory effects of CCT2 knockdown on proliferation and invasion. The findings suggest that CCT2 promotes HCC by activating the STAT3 signaling pathway. Therefore, CCT2 may serve as a survival biomarker and precision treatment target in HCC.
View Figures

Figure 1

In silico analysis of CCT2
expression and the clinical significance in HCC. Transcription
levels of CCT2 in (A) TCGA-LIHC, (B) ICGC-LIRI-JP and (C)
NODE-OEP00000321 datasets. ***P<0.001 vs. tumor-adjacent.
Association between CCT2 expression and the overall survival
prognosis of patients in (D) TCGA-LIHC, (E) ICGC-LIRI-JP and (F)
NODE-OEP00000321 datasets. CCT2, chaperonin containing TCP1 subunit
2; TCGA-LIHC, The Cancer Genome Atlas-liver hepatocellular
carcinoma; ICGC-LIRI-JP, International Cancer Genome
Consortium-liver Cancer-Riken, Japan; NODE, National Omics Data
Encyclopedia; HCC, hepatocellular carcinoma.

Figure 2

Knockdown of CCT2 in HCC cells. (A)
Protein expression levels of CCT2 in HCC cell lines (Huh-7, Hep3B,
Li-7 and HCCLM3). CCT2 was successfully knocked down in both Huh-7
and HCCLM3 cells following sh-CCT2 transduction. Knockdown
efficiency in Huh-7 cells was evaluated by (B) reverse
transcription-quantitative PCR and (C) western blotting. Knockdown
efficiency in HCCLM3 cells was evaluated by (D) reverse
transcription-quantitative PCR and (E) western blotting.
*P<0.05, ***P<0.001 vs. sh-NC. CCT2, chaperonin containing
TCP1 subunit 2; sh, short hairpin; NC, negative control; HCC,
hepatocellular carcinoma.

Figure 3

Knockdown of CCT2 inhibits the
proliferation and promotes the apoptosis of hepatocellular
carcinoma cells. EdU incorporation assay was used to determine the
DNA replication activity of (A) Huh-7 and (B) HCCLM3 cells. Colony
formation assay was used to assess the cell clonogenicity of (C)
Huh-7 and (D) HCCLM3 cells. Flow cytometry assay was used to
analyze the percentage of apoptotic (E) Huh-7 and (F) HCCLM3 cells.
Caspase-3/7 activity assay was used to evaluate the degree of
apoptosis in (G) Huh-7 and (H) HCCLM3 cells. *P<0.05,
**P<0.01, ***P<0.001 vs. sh-NC. CCT2, chaperonin containing
TCP1 subunit 2; sh, short hairpin; NC, negative control; RFU,
relative fluorescence units.

Figure 4

Knockdown of CCT2 inhibits the
migration and invasion of hepatocellular carcinoma cells. The
effect of CCT2 knockdown on the migration of (A) Huh-7 and (B)
HCCLM3 cells was measured using gap closure assay. The effects of
CCT2 knockdown on the invasion of (C) Huh-7 and (D) HCCLM3 cells
was evaluated by Transwell assay. *P<0.05, **P<0.01 vs.
sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short
hairpin; NC, negative control.

Figure 5

Knockdown of CCT2 inhibits the
stemness characteristics of hepatocellular carcinoma cells.
Tumor-sphere formation assay was used to assess the stem cell-like
properties of (A) Huh-7 and (B) HCCLM3 cells following CCT2
knockdown. EdU incorporation assay was used to determine the
proliferation of spheroid and parental (C) Huh-7 and (D) HCCLM3
cells following CCT2 knockdown. Transwell assay were used to
evaluate the invasion of spheroid and parental (E) Huh-7 and (F)
HCCLM3 cells following CCT2 knockdown. *P<0.05, **P<0.01,
***P<0.001 vs. parental sh-NC; #P<0.05,
##P<0.01 vs. spheroid sh-NC. CCT2, chaperonin
containing TCP1 subunit 2; sh, short hairpin; NC, negative control.
SFE, spheroid formation efficiency.

Figure 6

Knockdown of CCT2 inhibits the tumor
proliferation and metastasis of HCCLM3 cells in vivo. Tumor
(A) proliferation and (B) weights were measured to assess the
effects of CCT2 knockdown on HCCLM3 tumor proliferation in nude
mice. (C) Number of metastatic nodules stained with hematoxylin and
eosin was calculated to evaluate the effects of CCT2 knockdown on
the hematogenous lung metastasis of HCCLM3 cells (n=5/group).
*P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1
subunit 2; sh, short hairpin; NC, negative control.

Figure 7

Knockdown of CCT2 inhibits STAT3
signaling activation in hepatocellular carcinoma cells. The protein
levels of STAT3, p-STAT3, MCL1, MMP2 and SOX2 in (A) Huh-7 and (B)
HCCLM3 cells were measured by western blotting. The protein levels
of (C) p-STAT3, (D) MCL1, (E) MMP2 and (F) SOX2 in subcutaneous
tumor tissue were detected by immunohistochemical staining.
*P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1
subunit 2; sh, short hairpin; NC, negative control; p,
phosphorylated; MCL1, myeloid cell leukemia sequence 1; SOX2,
SRY-box transcription factor 2.

Figure 8

IL-6 abolishes the effects of CCT2
knockdown on the proliferation and invasion of hepatocellular
carcinoma cells. The protein levels of STAT3 and p-STAT3 in (A)
Huh-7 and (B) HCCLM3 cells were measured by western blotting. The
proliferation of (C) Huh-7 and (D) HCCLM3 cells following CCT2
knockdown and IL-6 treatment was detected using EdU incorporation
assay. The invasion of (E) Huh-7 and (F) HCCLM3 cells was assessed
using Transwell assay. *P<0.05, **P<0.01 vs. PBS. CCT2,
chaperonin containing TCP1 subunit 2; sh, short hairpin; p-,
phosphorylated.
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Li C, Yang L, Zhang Y, Zhao B and Qu H: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation. Oncol Rep 55: 81, 2026.
APA
Li, C., Yang, L., Zhang, Y., Zhao, B., & Qu, H. (2026). Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation. Oncology Reports, 55, 81. https://doi.org/10.3892/or.2026.9086
MLA
Li, C., Yang, L., Zhang, Y., Zhao, B., Qu, H."Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation". Oncology Reports 55.5 (2026): 81.
Chicago
Li, C., Yang, L., Zhang, Y., Zhao, B., Qu, H."Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation". Oncology Reports 55, no. 5 (2026): 81. https://doi.org/10.3892/or.2026.9086
Copy and paste a formatted citation
x
Spandidos Publications style
Li C, Yang L, Zhang Y, Zhao B and Qu H: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation. Oncol Rep 55: 81, 2026.
APA
Li, C., Yang, L., Zhang, Y., Zhao, B., & Qu, H. (2026). Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation. Oncology Reports, 55, 81. https://doi.org/10.3892/or.2026.9086
MLA
Li, C., Yang, L., Zhang, Y., Zhao, B., Qu, H."Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation". Oncology Reports 55.5 (2026): 81.
Chicago
Li, C., Yang, L., Zhang, Y., Zhao, B., Qu, H."Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation". Oncology Reports 55, no. 5 (2026): 81. https://doi.org/10.3892/or.2026.9086
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